纤维蛋白凝胶对上颌扩弓成骨矿化的影响

doi:10.19438/j.cjoms.2021.05.003

中国口腔颌面外科杂志 ›› 2021, Vol. 19 ›› Issue (5): 397-404.doi: 10.19438/j.cjoms.2021.05.003

纤维蛋白凝胶对上颌扩弓成骨矿化的影响

赵涵江¹, 王泽莹¹, 林丹^1,*, 沈国芳^1,2,*

1.上海交通大学医学院附属第九人民医院口腔颅颌面科,上海交通大学口腔医学院, 国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海 200011;
2.上海健康医学院,上海 201318

收稿日期:2021-03-18 修回日期:2021-05-11 出版日期:2021-09-20 发布日期:2021-10-20
通讯作者: 林丹,E-mail:d.lin@foxmail.com;沈国芳,E-mail:shengf@sumhs.edu.cn。^*共同通信作者
作者简介:赵涵江（1994-）,男,硕士研究生,E-mail:hanjiangzhao@sjtu.edu.cn
基金资助:
国家自然科学基金（81970973,81570947）; 上海市青年科技英才扬帆计划（19YF1426500）

Effect of fibrin glue on osteogenesis and mineralization during rapid maxillary expansion

ZHAO Han-jiang¹, WANG Ze-ying¹, LIN Dan¹, SHEN Guo-fang^1,2

1. Department of Oral and Craniomaxillofacial Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology. Shanghai 200011;
2. Shanghai University of Medicine and Health Sciences. Shanghai 201318, China

Received:2021-03-18 Revised:2021-05-11 Online:2021-09-20 Published:2021-10-20

摘要/Abstract

摘要： 目的研究原位注射纤维蛋白凝胶对SD大鼠上颌扩弓成骨矿化的影响。方法体外实验中,将大鼠骨髓间充质细胞（rBMSCs）在不同凝血酶浓度制备的纤维蛋白凝胶环境下进行共培养,通过CCK-8、 ALP染色和活性定量以及茜素红染色,检测梯度浓度凝血酶制备的纤维蛋白凝胶降解产物对rBMSCs细胞增殖和成骨分化的影响。将rBMSCs分别采用共培养、二维表面培养和三维培养方法与纤维蛋白凝胶进行结合,观察细胞形态和黏附情况。体内实验中,构建SD大鼠上颌扩弓模型（n=12）,以100 g作为扩弓力,扩弓7 d,随机分为2组,每组6只,将仅配戴14 d保持器作为对照组（R组）,将原位注射纤维蛋白凝胶联合配戴14 d保持器作为实验组（R+FG组）,通过显微CT（Micro-CT）三维重建和分析,比较新生骨量和骨矿化密度的差异。通过H-E染色、Masson三色染色、TRAP染色和顺序荧光染色,观察腭中缝的组织学差异。采用GraphPad Prism 8.0.1软件包对数据进行统计学分析。结果体外实验结果显示,纤维蛋白凝胶降解产物对rBMSCs细胞增殖和成骨分化无显著影响（P>0.05）,且以上述3种方式培养的rBMSCs均具有良好的细胞延展性。体内实验结果显示,实验组新生骨量（P<0.001）和骨小梁矿化密度（P<0.01）较对照组显著提升,腭中缝内可见大量矿化程度更高的新生骨小梁组织;破骨细胞数量减少,矿化沉积速率显著加快（P<0.001）。结论纤维蛋白凝胶降解产物对rBMSCs增殖、分化无影响。SD大鼠上颌扩弓后,腭中缝组织可通过原位注射纤维蛋白凝胶,促进新骨生成和矿化速率,抑制破骨细胞分化。

关键词: 上颌快速扩弓, 骨缝牵引成骨, 纤维蛋白凝胶, 骨再生

Abstract: PURPOSE: To investigate the effect of accelerating osteogenesis and mineralization during rapid maxillary expansion by in situ injection of fibrin glue. METHODS: In in vitro experiments, rat bone marrow stromal cells (rBMSCs) were co-cultured with fibrin glue fabricated with different concentrations of thrombin. The effects of degradation products of fibrin glue on the proliferation and osteogenic differentiation of rBMSCs were tested by CCK-8, alkaline phosphatase (ALP) staining and ALP activity quantification as well as alizarin red staining. rBMSCs were combined with fibrin glue by co-culture, two-dimensional surface culture and three-dimensional culture, respectively. Cell morphology and adhesion were observed by FITC-phalloidine fluorescence staining. In in vivo experiments, 12 6-week male SD rats were used to establish rapid maxillary expansion (RME) model with 100 g expanding force for 7 days. After 7-day expansion, they were divided into two groups: retention for 14 days as control group (R group) while in situ injection of fibrin glue combined with retention for 14 days as experimental group (R+FG group). Micro-CT scanning for three-dimensional reconstruction and analysis were conducted to compare the difference of new bone volume and bone mineral density. The osteogenesis and angiogenesis in mid-palatal suture were observed by H-E staining, Masson's trichrome staining, TRAP staining and sequential fluorescent labeling. GraphPad Prism 8.0.1 software package was used for statistical analysis. RESULTS: The degradation products of fibrin gel had little effect on cell proliferation and osteogenic differentiation of rBMSCs(P>0.05). In addition, rBMSCs cultured in three ways had great cell tractility. Compared with R group, the relative bone volume(P<0.001) and bone mineral density of trabeculae (P<0.001) were significantly increased in R+FG group. A large number of new trabeculae with higher mineralization degree were observed and the number of osteoclasts was decreased in mid-palatal suture. Mineral apposition rate was significantly accelerated in R+FG group(P<0.001). CONCLUSIONS: The degradation products of fibrin glue had no effect on proliferation and differentiation of rBMSCs. Fibrin glue could promote new bone formation, mineralized rate and inhibit osteo-clastogenesis in mid-palatal suture after RME in SD rats, which might be related to regulation of osteogenic differentiation by the surface structure of fibrin glue.

Key words: Rapid maxillary expansion, Sutural distraction osteogenesis, Fibrin glue, Bone regeneration

中图分类号:

R782.2

赵涵江, 王泽莹, 林丹, 沈国芳. 纤维蛋白凝胶对上颌扩弓成骨矿化的影响[J]. 中国口腔颌面外科杂志, 2021, 19(5): 397-404.

ZHAO Han-jiang, WANG Ze-ying, LIN Dan, SHEN Guo-fang. Effect of fibrin glue on osteogenesis and mineralization during rapid maxillary expansion[J]. China J Oral Maxillofac Surg, 2021, 19(5): 397-404.

参考文献

[1] Zandi M, Miresmaeili A, Heidari A.Short-term skeletal and dental changes following bone-borne versus tooth-borne surgically assisted rapid maxillary expansion: a randomized clinical trial study[J]. J Craniomaxillofac Surg, 2014, 42(7): 1190-1195.
[2] Carlson C, Sung J, McComb RW, et al. Microimplant-assisted rapid palatal expansion appliance to orthopedically correct transverse maxillary deficiency in an adult[J]. Am J Orthod Dentofacial Orthop, 2016, 149(5): 716-728.
[3] Neyt NM, Mommaerts MY, Abeloos JV, et al.Problems, obstacles and complications with transpalatal distraction in non-congenital deformities[J]. J Craniomaxillofac Surg, 2002, 30(3): 139-143.
[4] Vandersea BA, Ruvo AT, Frost DE.Maxillary transverse deficiency - surgical alternatives to management[J]. Oral Maxillofac Surg Clin North Am, 2007, 19(3): 351-368.
[5] Khosravi M, Ugolini A, Miresmaeili A, et al.Tooth-borne versus bone-borne rapid maxillary expansion for transverse maxillary deficiency: a systematic review[J]. Int Orthod, 2019, 17(3): 425-436.
[6] Fatima F, Jeelani W, Ahmed MJD, et al.Current trends in craniofacial distraction: a literature review[J]. Dent Med Probl, 2020, 57(4): 441-448.
[7] Andrucioli MCD, Matsumoto MAN.Transverse maxillary deficiency: treatment alternatives in face of early skeletal maturation[J]. Dental Press J Orthod, 2020, 25(1): 70-79.
[8] Aloise AC, Pereira MD, Hino CT, et al.Stability of the transverse dimension of the maxilla after surgically assisted rapid expansion[J]. J Craniofac Surg, 2007, 18(4): 860-865.
[9] Moussa R, O'Reilly MT, Close JM. Long-term stability of rapid palatal expander treatment and edgewise mechanotherapy[J]. Am J Orthod Dentofacial Orthop, 1995, 108(5): 478-488.
[10] Ramstad T, Jendal T.A long-term study of transverse stability of maxillary teeth in patients with unilateral complete cleft lip and palate[J]. J Oral Rehabil, 1997, 24(9): 658-665.
[11] Dreifke MB, Ebraheim NA, Jayasuriya AC.Investigation of potential injectable polymeric biomaterials for bone regeneration[J]. J Biomed Mater Res A, 2013, 101(8): 2436-2447.
[12] Tamimi F, Torres J, Lopez-Cabarcos E, et al.Minimally invasive maxillofacial vertical bone augmentation using brushite based cements[J]. Biomaterials, 2009, 30(2): 208-216.
[13] Zhou Y, Ni Y, Liu Y, et al.The role of simvastatin in the osteogenesis of injectable tissue-engineered bone based on human adipose-derived stromal cells and platelet-rich plasma[J]. Biomaterials, 2010, 31(20): 5325-5335.
[14] Ingavle GC, Gionet-Gonzales M, Vorwald CE, et al.Injectable mineralized microsphere-loaded composite hydrogels for bone repair in a sheep bone defect model[J]. Biomaterials, 2019, 197: 119-128.
[15] Terbish M, Yoo SH, Kim HJ, et al.Accelerated bone formation in distracted alveolar bone after injection of recombinant human bone morphogenetic protein-2[J]. J Periodontol, 2015, 86(9): 1078-1086.
[16] Wang L, Zhou S, Liu B, et al.Locally applied nerve growth factor enhances bone consolidation in a rabbit model of mandibular distraction osteogenesis[J]. J Orthop Res, 2006, 24(12): 2238-2245.
[17] Cho BC, Chung HY, Lee DG, et al.The effect of chitosan bead encapsulating calcium sulfate as an injectable bone substitute on consolidation in the mandibular distraction osteogenesis of a dog model[J]. J Oral Maxillofac Surg, 2005, 63(12): 1753-1764.
[18] Wolberg AS.Thrombin generation and fibrin clot structure[J]. Blood Rev, 2007, 21(3): 131-142.
[19] Adam SS, Key NS, Greenberg CS.D-dimer antigen: current concepts and future prospects[J]. Blood, 2009, 113(13): 2878-2887.
[20] Padilla S, Sanchez M, Orive G, et al.Human-based biological and biomimetic autologous therapies for musculoskeletal tissue regeneration[J]. Trends Biotechnol, 2017, 35(3): 192-202.
[21] Noori A, Ashrafi SJ, Vaez-Ghaemi R, et al.A review of fibrin and fibrin composites for bone tissue engineering[J]. Int J Nanomedicine, 2017, 12: 4937-4961.
[22] McDuffee LA, Esparza Gonzalez BP, Nino-Fong R, et al. Evaluation of an in vivo heterotopic model of osteogenic differentiation of equine bone marrow and muscle mesenchymal stem cells in fibrin glue scaffold[J]. Cell Tissue Res, 2014, 355(2): 327-335.
[23] Abiraman S, Varma HK, Umashankar PR, et al.Fibrin glue as an osteoinductive protein in a mouse model[J]. Biomaterials, 2002, 23(14): 3023-3031.
[24] Wang L, Zhang C, Li C, et al.Injectable calcium phosphate with hydrogel fibers encapsulating induced pluripotent, dental pulp and bone marrow stem cells for bone repair[J]. Mater Sci Eng C Mater Biol Appl, 2016, 69: 1125-1136.
[25] Dong J, Cui G, Bi L, et al.The mechanical and biological studies of calcium phosphate cement-fibrin glue for bone reconstruction of rabbit femoral defects[J]. Int J Nanomedicine, 2013, 8: 1317-1324.
[26] van Esterik FA, Zandieh-Doulabi B, Kleverlaan CJ, et al. Enhanced osteogenic and vasculogenic differentiation potential of human adipose stem cells on biphasic calcium phosphate scaffolds in fibrin gels[J]. Stem Cells Int, 2016, 2016: 1934270.
[27] Oh JH, Kim HJ, Kim TI, et al.The effects of the modulation of the fibronectin-binding capacity of fibrin by thrombin on osteoblast differentiation[J]. Biomaterials, 2012, 33(16): 4089-4099.
[28] Arkudas A, Pryymachuk G, Hoereth T, et al.Composition of fibrin glues significantly influences axial vascularization and degradation in isolation chamber model[J]. Blood Coagul Fibrinolysis, 2012, 23(5): 419-427.
[29] Hong C, Song D, Lee DK, et al.Reducing posttreatment relapse in cleft lip palatal expansion using an injectable estrogen-nanodiamond hydrogel[J]. Proc Natl Acad Sci USA, 2017, 114(35): E7218-E7225.
[30] Segura-Castillo JL, Aguirre-Camacho H, González-Ojeda A, et al.Reduction of bone resorption by the application of fibrin glue in the reconstruction of the alveolar cleft[J]. J Craniofac Surg, 2005, 16(1): 105-112.
[31] Giannini G, Mauro V, Agostino T, et al.Use of autologous fibrin-platelet glue and bone fragments in maxillofacial surgery[J]. Transfus Apher Sci, 2004, 30(2): 139-144.
[32] Pagel C, Song S-J, Loh L, et al.Thrombin-stimulated growth factor and cytokine expression in osteoblasts is mediated by protease-activated receptor-1 and prostanoids[J]. Bone, 2009, 44(5): 813-821.
[33] Nair M, Varma HK, John A.Platelet-rich plasma and fibrin glue-coated bioactive ceramics enhance growth and differentiation of goat bone marrow-derived stem cells[J]. Tissue Eng Part A, 2009, 15(7): 1619-1631.
[34] 刘红, 杨超, 陈国庆, 等. 纤维蛋白胶不同复合方式对牙囊细胞增殖活性的影响[J]. 华西口腔医学杂志, 2015, 33(2): 135-140.
[35] Roberts IV, Bukhary D, Valdivieso CYL, et al.Fibrin matrices as (injectable) biomaterials: formation, clinical use, and molecular engineering[J]. Macromol Biosci, 2020, 20(1): e1900283.
[36] Tang GH, Xu J, Chen RJ, et al.Lithium delivery enhances bone growth during midpalatal expansion[J]. J Dent Res, 2011, 90(3): 336-340.
[37] Altan AB, Bicakci AA, Avunduk MC, et al.The effect of dosage on the efficiency of LLLT in new bone formation at the expanded suture in rats[J]. Lasers Med Sci, 2015, 30(1): 255-262.
[38] Kara MI, Erciyas K, Altan AB, et al.Thymoquinone accelerates new bone formation in the rapid maxillary expansion procedure[J]. Arch Oral Biol, 2012, 57(4): 357-363.
[39] Yuan Z, Wei P, Huang Y, et al.Injectable PLGA microspheres with tunable magnesium ion release for promoting bone regeneration[J]. Acta Biomater, 2019, 85: 294-309.

纤维蛋白凝胶对上颌扩弓成骨矿化的影响

Effect of fibrin glue on osteogenesis and mineralization during rapid maxillary expansion

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	王博, 于洪波, 夏韫晖, 房兵, 郭秋曼. 牙槽骨再生正畸技术治疗成人错畸形伴原发性牙槽骨缺损的远期疗效评价[J]. 中国口腔颌面外科杂志, 2022, 20(3): 225-229.
[2]	唐田, 程群, 丁子清, 朱信灵, 胡英英. MTA与PRF在上颌前牙拔牙患者牙槽骨再生中的应用效果评价[J]. 中国口腔颌面外科杂志, 2022, 20(3): 263-267.
[3]	李晶, 梅东梅, 姜雅萍, 徐昊, 王文雪, 姚晨阳, 李亚男, 赵保东. 2 mm矮愈合基台在前牙美学区早期种植骨增量中的效果分析[J]. 中国口腔颌面外科杂志, 2021, 19(6): 531-535.
[4]	费晨艳, 江银华. 生长因子在牙槽骨组织再生中的应用与研究进展[J]. 中国口腔颌面外科杂志, 2021, 19(6): 568-572.
[5]	邹多宏, 刘昌奎, 薛洋, 胡开进, 杨驰, 张志愿. 帐篷钉技术在牙槽骨修复与再生中的临床应用及操作规范[J]. 中国口腔颌面外科杂志, 2021, 19(1): 1-5.
[6]	李成, 施乐. 2种修复膜材料用于牙种植引导骨再生的临床效果比较[J]. 中国口腔颌面外科杂志, 2020, 18(5): 438-441.
[7]	王明一, 傅振, 张陈平. 血管化腓骨重建下颌骨后用腓骨残端游离移植及牙种植的可行性研究[J]. 中国口腔颌面外科杂志, 2020, 18(4): 338-342.
[8]	秦坤, 刘红红, 章润宇, 张志宏. 引导骨再生术对前牙区种植牙龈美学及牙槽骨吸收的影响[J]. 中国口腔颌面外科杂志, 2020, 18(3): 236-239.
[9]	姚志涛, 安玮, 买买提吐逊·吐尔地. 50例唇腭裂患儿手术前、后血液生化指标及腭裂隙内骨再生评价[J]. 中国口腔颌面外科杂志, 2020, 18(1): 56-59.
[10]	姜帅, 姜雅萍, 李晓静, 王文雪, 滕敏华, 李欣, 赵保东, 梅东梅. 去膜化GBR技术在后牙种植中的应用[J]. 中国口腔颌面外科杂志, 2019, 17(3): 251-256.
[11]	聂萍，陶丽，孙蕙珺，于倩，朱敏. 手术辅助上颌骨快速扩弓对鼻气道流场特征的影响[J]. 中国口腔颌面外科杂志, 2018, 16(3): 237-241.
[12]	张楚南, 倪杰, 莫嘉骥, 乔士冲, 王蓓, 顾迎新. 美学区种植同期应用异种骨行引导骨再生术后愈合期间唇侧骨改建的临床研究[J]. 中国口腔颌面外科杂志, 2018, 16(1): 29-33.
[13]	戈旌, 杨驰, 郑家伟, 汪湧, 华洪飞. 阻生第三磨牙拔除术后自体骨回植第二磨牙远中骨缺损的单中心随机对照临床研究[J]. 中国口腔颌面外科杂志, 2017, 15(4): 335-340.
[14]	李慧萍, 孙守福, 甄锦泽, 王烨欣, 徐伟峰, 郑吉驷, 张善勇, 朱应超, 钟晓琪. 钛网+膜联合使用修复种植体周围牙槽骨缺损的实验研究[J]. 中国口腔颌面外科杂志, 2017, 15(2): 109-114.
[15]	李慧萍, 孙守福, 甄锦泽, 王烨欣, 徐伟峰, 郑吉驷, 张善勇, 朱应超, 钟晓琪. 钛网+膜联合使用修复种植体周围牙槽骨缺损的实验研究[J]. 中国口腔颌面外科杂志, 2017, 15(2): 127-130.